| Objective To study the positive rate of HBsAg^anti-HBs and the immune non-response status of hepatitis B vaccine and the factors effecting hepatitis B immune response failure among Primary and middle school students. To investigate the positive rate of HBV DNA and S gene mutations and select the evolutionary model based on the S gene fragment. To investigate the genotypes of hepatitis B virus and analysis of Amino acid variation.And provide a scientific basis for hepatitis B prevention and control. Method Adopt Hepatitis B serological test of three primary and middle school students in Urumqi County. Detected two serological markers of hepatitis B HBsAg and anti-HBs by ELISA method. The469students who the negative of HBsAg and anti-HBs were injected two vaccine immunization coverage. The anti-HBs was detected by ELISA in the second vaccination one month again. The anti-HBs no response group (case group) and response group (control group) were found according to the result of the detection and to investigate the related factors. The1:2case-control of Single factor analysis and multiple factor logistic analysis were performed by SPSS17.0software. The469venous blood samples that the two serological markers of HBsAg and anti-HBs were negative by ELISA detection for finding HBV DNA with trace DNA extraction kit. Using the polymerase chain reaction (PCR) amplified of HBV DNA gene fragments, analyzing the infection rate of occult hepatitis B virus (OBI). The10cases of positive samples were PCR amplified sequence and founded a HBV sequence database, and compared and analyzed to get the hepatitis B virus S gene fragment and the homologous relationship and nucleotide/amino acid variation between the sequencing results and the NCBI international nucleotide sequence database collaborative group (INSDC, Genebank) by DNA MEGA5.1software. Constructed the evolutionary tree and select the evolutionary model based on comparing S gene difference between determination of reference sequences and ones in INSDC included to investigate genotype determination of virus and the genotype distribution difference and the nucleotide variation. Result Primary and middle school students before immunization positive rate of HBsAg was1.46%. The HBsAg positive rate of primary school students was lower than middle and no statistically significant difference between male and female. There was difference among Han, Hui, Kazak, Uygur and other ethnic groups.The study of hepatitis B infection after the hepatitis B vaccine immunization of primary and middle school students. The test showed that sero-conversion rate is88.06%in a month for two injections(0and1th month) of hepatitis B vaccine for groups of Antigen-antibody were negative.No/Weak response rate was11.94%and no significant difference between primary and middle school students (P>0.05).Univariate analysis indicated the factors had associated with non-responsive-ness, such as exelcymosis (OR=4.932,95%IC1.091～22.295), neonatal birth weight (OR=0.255,95%IC0.116～0.560), delivery place (OR=0.361,95%IC0.165～0.790), feedway (OR=0.423,95%CI0.179～0.999), neonatus vaccination (OR=0.453,95%CI0.209～0.980).The results of multivariate logistic regression showed that only four factors were entered in the model including exelcymosis (OR=5.416,95%CI1.197～24.506), neonatal birth weight (OR=0.410,95%CI0.172～0.979), delivery place (OR=0.445,95%CI0.213～0.929), neonatus vaccination (OR=0.399,95%CI0.184～0.863.The469serum samples were found43HBV positive samples by PCR amplification and sequenced DNA.10for the complete sequencing in this papers,4copies of the gene mutations. Among them,1case of variable sites in alpha determinant in the range of124-147amino acids.1case of variable sites in alpha determinant upstream72amino acid insert C frameshift to mutations of amino acids.2cases of variable sites concentrated in alpha determinant downstream.Conclusion Primary and middle school students anti-HBs positive rate was relatively low.Hepatitis B vaccination is necessary for students. Many factors were associated with non-responsiveness of hepatitis B vaccine. Extraction is the dangerous factors of vaccine no response. Neonatal normal birth weight and neonatal vaccination within 24hours were protective factors promote immune response.The mutation of S gene of hepatitis B vaccine immunization in different sites were formed among primary and middle school students after inoculation of hepatitis B, the causes of immunity failure may be caused by the hepatitis B vaccine and; no mutant strains has little effect hepatitis B vaccine on the current... |
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